Detaljerad miljöinformation

Environmental Risk Classification

Predicted Environmental Concentration (PEC)

PEC is calculated according to the following formula: (Ref.1)

PEC (μg/L) = (A*10⁹*(100-R))/(365*P*V*D*100)

Where:

A (kg/yr)	0,13 kg	Total glekaprevir (ABT-493) sold (kg) in Sweden in 2020 from IQVIA (Ref. 2)
R	0 %	Removal rate (due to loss by adsorption to sludge particles, by volatilization, hydrolysis or biodegradation); use 0 if no data is available. (Ref.1)
P	10*106	Number of inhabitants in Sweden (Ref. 1)
V (L/day)	200	Volume of wastewater per capita and day (200 L/day is the default value) (Ref. 1,3)
D	10	Factor for dilution of wastewater by surface water flow (10 is the default value) (Ref. 1,3).

Note: The factor 10⁹ converts the quantity distressed from kg to mcg.

PEC (μg/L) = (0,13*10⁹*(100-0))/(365*10*10⁶*200*10*100)

PEC = 0,000018 μg/L

Ecotoxicological Studies with Glekaprevir

Activated Sludge, Respiration Inhibition Test (OECD 209)

The potential effect of glekaprevir on the respiration rate of activated sludge microorganisms was investigated according to OECD Guideline 209. (Ref. 4)

Endpoint	Time	Result (mg/L)
EC50	30 Min	> 1000
EC50	3 Hr	> 1000

The respiration rate of the 1000 mg glekaprevir treatment in the limit test exceeded the control treatment. Therefore, the NOEC is 1000 mg/L.

Freshwater Alga Growth Inhibition Test (OECD 201)

The effect of glekaprevir on the yield and average specific growth rate of the freshwater green alga, Pseudokirchneriella subcapitata, was determined in accordance with OECD Guideline 201. (Ref. 5)

	Effect Concentration (mg glekaprevir/L)a
	72-Hour Yield	72-Hour Average Specific Growth Rate
NOEC	16,9	40,6

^a.Expressed as geometric mean measured concentration of glekaprevir, (mg/L).

The most conservative NOEC was 16,9 mg/L (algal yield).

Daphnia magna Reproduction Test (OECD 211)

The potential for chronic effects of glekaprevir on survival, reproduction, and growth of the freshwater crustacean, Daphnia magna was investigated in accordance with OECD Guideline 211. (Ref. 6) The results are below.

	Effect Concentration (mg glekaprevir/L)a,b
	Survival	Reproduction			Growth
		Young/Adult Reproductive Day	Young/Adult at Study Initiation	Young/Adult at Test Termination	Length
21-day NOEC	4,91	4,91	4,91	4,91	4,91

^a.Replicate C of 0,250 mg/L and Replicate A of 0,562 mg/L excluded from statistical analyses because survival in these replicates were shown to be statistical outliers.

^b.Expressed as arithmetic mean measured concentration of glekaprevir, (mg/L).

The NOEC was 4,91 mg/L for all survival, reproduction, and growth endpoints.

Fish Early-Life Stage Toxicity Test (OECD 210)

The chronic effects of glekaprevir to the fathead minnow, Pimephales promelas, embryos and larvae under flow-through conditions were tested in accordance with OECD 210, with the following results. (Ref. 7)

	Effect Concentration (mg glekaprevir/L)a
	Egg Hatch, Time to Hatch Start and Hatch End	Fry Survivalb	Total Lengthc	Blotted Weight
NOEC	10,2	10,2	10,2	10,2

^a. Expressed as mean measured concentration of glekaprevir, (mg/L).

^b. Fry survival based on number of hatched fry surviving on Day 28 post-hatch.

^c. A statistically significant reduction at the 4,89 mg/L test treatment was not considered to be biologically significant because the reduction was only 2% as compared to the pooled control and there was no statistically signification reduction at the 10,2 mg/L test treatment

The NOEC was determined to be 10,2 mg/L for all survival, hatch, and growth endpoints.

Predicted No Effect Concentration (PNEC)

PNEC (μg/L) = NOEC/AF

AF = Assessment Factor = 10

Organism	NOEC
Microorganisms (spps)	1000 mg/L
Freshwater Algae (Pseudokirchneriella subcapitata)	40,6 mg/L (average specific growth rate)
	16,9 mg/L (algal yield)
Daphnia magna	4,91 mg/L (all endpoints)
Fathead Minnow (Lepomis macrochirus)	10,2 mg/L (all endpoints)

The PNEC was determined in accordance with ECHA guidance (Ref. 8).

The chronic aquatic effects of glekaprevir were assessed in green algae, fish, and Daphnia. Daphnia magna was determined to be the most sensitive species tested (NOEC of 4,91 mg/L). Therefore, the PNEC_SURFACEWATER was calculated using the NOEC for Daphnia magna.

NOEC = 4,91 mg/L

PNEC (mg/L) = 4,91/10

PNEC = 0,491 mg/L

PNEC = 491 μg/L

Environmental Risk Classification (PEC/PNEC ratio)

PEC/PNEC Ratio:

PEC = 0,000018 μg/L

PNEC = 491 μg/L

PEC/PNEC = 0,000018/491

PEC/PNEC = 0,000000036

Justification of environmental risk classification:

Since PEC/PNEC ≤ 0,1, the use of glekaprevir has been considered to result in insignificant environmental risk.

Degradation

Aerobic Transformation in Aquatic Sediment Systems (OECD 308)

The transformation of [¹⁴C]glekaprevir was investigated according to OECD Guideline 308, in two different water/sediment systems under aerobic conditions. (Ref. 9)

The degradation and distribution of [¹⁴C]ABT-493 was investigated according to OECD Guideline 308. The transformation of [¹⁴C]ABT-493 was studied in two different water/sediment systems (Brandywine River and Choptank River) under aerobic conditions.

According to the study report, the water phase of the test system was treated with

[¹⁴C]ABT-493 at a rate of 0,5 μg/mL water and incubated in darkness at approximately 20 ± 2°C for up to 100 days. Aerobic conditions were maintained by passing a steady stream of humidified air through the test apparatus. The flow-through systems trapped evolved carbon dioxide (CO2) and volatile organic compounds. Duplicate samples from each radiolabelled treatment were taken at 0, 3, 7, 14, 30, 50, 70, and 100 days after application. The water and sediment layers were separated by centrifugation

and decanting. The water layer was analyzed by liquid scintillation counting (LSC) for

radioactivity content and then analyzed by direct injection using reversed-phase high performance liquid chromatography (HPLC) to determine the biotransformation products. The sediment layer was extracted with appropriate organic solvent and were analyzed by LSC, concentrated, filtered, and analyzed by reverse-phase HPLC. The post-extracted sediment pellet was homogenized, combusted, and the radioactive residue quantified by LSC. Volatiles traps (2 N KOH) were analyzed by LSC.

The amount of radioactivity in the sediment layer (extractable residue) generally increased over the course of the study from a mean of 19,5% applied radioactivity (AR) at Day 0 to a mean of 59,7% AR at Day 100 for the Brandywine sediments and from a mean of 28,7% AR at Day 0 to a mean of 53,7% AR at Day 100 for the Choptank sediments (the lowest mean for Choptank sediments (22%) was measured on Day 3).Non-extractable residues increased steadily over the duration of the study, reaching a mean maximum of 38,9 and 22,3% AR at Days 100 and 70 for the Brandywine and Choptank samples, respectively.

The half-lives for water, sediment, and total system were determined to be as follows:

	Layer	Brandywine River	Choptank River
DT50, 20°C (days)	Water	25,5	39,5
	Sediment	253	284
	Total System	42,1	52,5

As is shown, the DT₅₀ values for the total system in the Brandywine and Choptank Rivers were 42,1 and 52,5 days, respectively.

Justification of chosen degradation phrase:

DT₅₀< 120d for the total system; therefore, glekaprevir is slowly degraded in the environment.

Bioaccumulation

Partition Coefficient (n-octanol/water): Shake Flask Method

The n-octanol-buffer distribution coefficient of glekaprevir was determined using a shake flask method at pH 7,4 and 25°C. (Ref. 10,11)

Endpoint	Result	pH
Log D	2,50 ± 0,0	7,4

Justification of chosen bioaccumulation phrase:

Log D_ow at pH 7< 4,0; therefore, glekaprevir has low potential for bioaccumulation.

References

1. FASS.se. Environmental classification of pharmaceuticals at www.fass.se. Guidance for pharmaceutical companies. 2012 V 2.0. 2021.

2. IQVIA. 2020. IQVIA / LIF - kg consumption/2020.

3. European Chemicals Agency (ECHA). Guidance on Information Requirements and Chemical Safety Assessment Chapter R.16: Environmental exposure assessment. Version 3.0. 2016.

4. EAG. R&D/16/0263. ABT-493: Activated Sludge, Respiration Inhibition Test. ABC Study 82939. AbbVie Study TX15-194. Amended 09 January 2017.

5. EAG. R&D/16/0260. ABT-493: Growth Inhibition Test with the Unicellular Green Alga, Pseudokirchneriella subcapitata. ABC Study 82936. AbbVie Study TX15-191. Amended 08 June 2017.

6. EAG. R&D/16/0261. ABT-493: Chronic Toxicity Test with the Cladoceran, Daphnia magna, Conducted Under Flow-Through Conditions. ABC Study 82938. AbbVie Study TX15-192. 09 December 2016.

7. EAG. R&D/16/0262. ABT-493: Early Life-Stage Toxicity Test with the Fathead Minnow, Pimephales promelas, Under Flow-Through Conditions. ABC Study 82937. AbbVie Study TX15-193. 08 December 2016.

8. European Chemicals Agency (ECHA). Guidance on information requirements and chemical safety assessment Chapter R.10: Characterisation of dose [concentration]-response for environment. 2008.

9. EAG. R&D/16/0744. [14C]ABT-493: Aerobic Aquatic Metabolism in Two Sediment Systems. EAG Study 82942. AbbVie Study TX15-189. Amended 21 June 2017.

10. AbbVie. PTR-16-0010. Physicochemical Properties of ABT-493 (Glecaprevir, A 1282576). 2016.

11. AbbVie. PME-17-0001. Preformulation Memo (PME): A-1282576.0 (ABT-493) Distribution Coefficient (Log D). 2017.

Detaljerad miljöinformation

Environmental Risk Classification

Predicted Environmental Concentration (PEC)

PEC is calculated according to the following formula: (Ref.1)

PEC (μg/L) = (A*10⁹*(100-R))/(365*P*V*D*100)

Where:

A (kg/yr)	0,04 kg	Total pibrentasvir (ABT-530) sold (kg) in Sweden in 2020 from IQVIA (Ref. 2)
R	0 %	Removal rate (due to loss by adsorption to sludge particles, by volatilization, hydrolysis or biodegradation); use 0 if no data is available. (Ref.1)
P	10*106	Number of inhabitants in Sweden (Ref. 1)
V (L/day)	200	Volume of wastewater per capita and day (200 L/day is the default value) (Ref. 1;3)
D	10	Factor for dilution of wastewater by surface water flow (10 is the default value). (Ref. 1;3)

Note: The factor 10⁹ converts the quantity distressed from kg to mcg.

PEC (μg/L) = (0,04 *10⁹*(100-0))/(365*10*10⁶*200*10*100)

PEC = 0,000005 μg/L

Ecotoxicological Studies with Pibrentasvir

Activated Sludge, Respiration Inhibition Test (OECD 209)

The effect of pibrentasvir on the respiration rate of activated sludge was assessed using the methods that met the requirements of OECD 209 and OECD 54 with the following results. (Ref. 4)

Endpoint	Time	Result (mg/L)
EC20	3 Hr	> 1000
EC50	3 Hr	> 1000
EC80	3 Hr	> 1000

Pibrentasvir did not significantly inhibit the respiration rates of the samples of activated sludge, with a maximum reduction in respiration of 9% at 1000 mg/L, which was not statistically significant. The no observed effect concentration (NOEC) for pibrentasvir was 1000 mg/L (the highest concentration tested).

Freshwater Alga Growth Inhibition Test (OECD 201)

A study was performed to assess the toxicity of pibrentasvir on the growth of the green alga Pseudokirchneriella subcapitata. The method complied with the OECD 201; the results are shown below. (Ref. 5)

	Effect Concentration (mg pibrentasvir/L)a
	Inhibition of growth rate	Inhibition of Yield	Inhibition of Biomass Integral
NOEC	0,0044	0,0044	0,0044

^a.Based on the geometric mean measured test concentration.

There were no statistically significant decreases in growth rate, yield, and/or biomass. Therefore, the NOEC was 0,0044 mg/L.

Daphnia magna Reproduction Test (OECD 211)

A study was performed to assess the chronic toxicity of pibrentasvir to the freshwater crustacean, Daphnia magna (also known as water fleas) following OECD 211. (Ref. 6) The results were as follows.

	Effect Concentration (mg pibrentasvir/L)a
	Survival	Reproduction	Growth
21-day NOEC	0,0057	0,0057	0,0057

^a.Based on the geometric mean measured test concentrations.

The NOEC was 0,0057 mg/L for all survival, reproduction, and growth endpoints.

Fish Early-Life Stage Toxicity Test (OECD 210)

A study was performed to assess the effects of pibrentasvir on freshly hatched larvae of the fathead minnow (Pimephales promelas) under flow-through conditions in accordance with OECD 210. (Ref. 7) The study yielded the following results.

	Effect Concentration (mg pibrentasvir/L)a
	Number of Eggs Hatching	Post-Hatch Survival	Length	Weight
NOEC	0,003	0,003	0,003	0,003

a. Expressed as measured concentration of pibrentasvir, (mg pibrentasvir /L).

The NOEC was determined to be 0,0030 mg/L for all survival, hatch, and growth endpoints. This study showed that there were no toxic effects at the limit of solubility.

Predicted No Effect Concentration (PNEC)

PNEC (μg/L) = NOEC/AF

AF = Assessment Factor= 10

Organism	NOEC
Microorganisms (spps)	1000 mg/L
Freshwater Algae (Pseudokirchneriella subcapitata)	0,0044 mg/L (all endpoints)
Daphnia magna	0,0057 mg/L (all endpoints)
Fathead Minnow (Lepomis macrochirus)	0,003 mg/L (all endpoints)

The PNEC was determined in accordance with ECHA guidance (Ref. 8).

The chronic aquatic effects of pibrentasvir were assessed in green algae, fish, and Daphnia. The fathead minnow (Lepomis macrochirus) was determined to be the most sensitive species tested (NOEC of 0,0030 mg/L). Therefore, the PNEC was calculated using the NOEC for the fathead minnow.

NOEC = 0,003 mg/L

PNEC (mg/L) = 0,003/10

PNEC = 0,0003 mg/L

PNEC = 0,30 μg/L

Environmental Risk Classification (PEC/PNEC ratio)

PEC/PNEC Ratio:

PEC = 0,000005 μg/L

PNEC = 0,30 μg/L

PEC/PNEC = 0,000005/0,30

PEC/PNEC = 0,0002

Justification of environmental risk classification:

Since PEC/PNEC ≤ 0,1; therefore, the use of pibrentasvir has been considered to result in insignificant environmental risk.

Degradation

Aerobic Transformation in Aquatic Sediment Systems (OECD 308)

The degradation and distribution of [¹⁴C]pibrentasvir was investigated according to OECD Guideline 308, aerobic transformation in aquatic sediment systems. (Ref. 9)  The fate of pibrentasvir was studied under (aerobic) laboratory conditions using two natural aquatic sediment systems from Swiss Lake and Calwich Abbey Lake.

According to the study report, the aquatic sediment samples were treated with [¹⁴C]pibrentasvir at a rate of 0,45 mg/L and incubated under aerobic conditions at 20 ± 2°C in darkness for periods of up to 100 days.  Aerobic conditions were maintained by passing a steady stream of humidified air through the test apparatus.

Total recoveries of radioactivity (mass balances) were between 65,9% and 118,2% applied radioactivity. It was determined that the low mass balances were due to binding of the test item to glassware used.

In Calwich Abbey Lake aquatic sediment, the total radioactivity in the water layer declined quickly to 5,8 – 6,8% applied radioactivity by 7 days and then remained at a similar level during the rest of the incubation period. In sediment, the total radioactivity increased to 88,6 – 90,0% applied radioactivity after 7 days and remained at a similar level up to 100 days of incubation. The proportion of radioactivity remaining unextracted in the sediment increased to 5,7 – 6,0% of applied radioactivity after 100 days.

Dissipation of radioactivity in the Swiss Lake aquatic sediment was faster than for Calwich Abbey. The total radioactivity in the water layer declined to 15,4 – 23,8% after 7 days and decreased to 14,2 – 17,9% after 100 days. In sediment, the total radioactivity increased to 68,1 – 83,3% applied radioactivity after 7 days and increased to 82,5 – 86,1% applied radioactivity after 100 days of incubation. The proportion of radioactivity remaining unextracted in the sediment increased to 29,0 – 40,5% of applied radioactivity after 30 days.

Pibrentasvir dissipated rapidly from the water of aquatic sediment systems.  The half-lives of pibrentasvir (parent compound only) for water, sediment, and total system were determined to be as follows:

	Layer	Calwich Abbey Lake	Swiss Lake
DT50 (days, 20°C)	Water	1,9	1,2
	Sediment	956	38
	Total system	395	6,5

As is shown, the DT₅₀ values for the total system in Calwich Abbey Lake and Swiss Lake were 395 and 6,5 days, respectively.

Justification of chosen degradation phrase:

As the highest DT₅₀ > 120d for the total system, pibrentasvir is potentially persistent.

Bioaccumulation

Partition Coefficient (n-octanol/water): Shake Flask Method

The n-octanol-buffer distribution coefficient was determined using a shake flask method at pH 7,4 and 25°C. (Ref. 10)

Endpoint	Result	pH
Log D	7,47 ± 0,03	7,4

Justification of chosen bioaccumulation phrase:

CLP (classification, labelling and packaging) guidance states that a Log Kow (Log D) > 4 indicates the substance may bioaccumulate. (Ref. 1) The Log D of pibrentasvir is 7,47.  Therefore, pibrentasvir has high potential for bioaccumulation.

References

1. FASS.se. Environmental classification of pharmaceuticals at www.fass.se. Guidance for pharmaceutical companies. 2012 V 2,0. 2021.

2. IQVIA. 2020. IQVIA / LIF - kg consumption/2020.

3. European Chemicals Agency (ECHA). Guidance on Information Requirements and Chemical Safety Assessment Chapter R.16: Environmental exposure assessment. Version 3.0. 2016.

4. Envigo. R&D/16/0982. ABT-530: Activated Sludge Respiration Inhibition Test. Envigo Study Number: HF35KP. 15 August 2016. Amended 04 October 2017.

5. Envigo. R&D/16/1081. ABT-530: Algal Growth Inhibition Test (OECD 201). Envigo Study Number: MG48TK. 04 May 2017. Amended 21 September 2018.

6. Envigo. R&D/16/1083. ABT-530: Daphnia magna Reproduction Test. Envigo Study Number: HB89KG. 12 June 2017. Amended 16 October 2017.

7. Envigo. R&D/16/1082. ABT-530: Fish, Early Life Stage Toxicity Test. Envigo Study Number: HV85WK. 12 January 2018. Amended 28 September 2018.

8. European Chemicals Agency (ECHA). Guidance on information requirements and chemical safety assessment Chapter R.10: Characterisation of dose [concentration]-response for environment. 2008.

9. Envigo. R&D/16/1085. ABT-530: Aerobic Transformation in Aquatic Sediment Systems. Envigo Study Number: YY54JC. 18 July 2017.

10. AbbVie. PTR-16-0009. Physicochemical Properties of ABT-530 (Pibrentasvir, A 1325912). 02 November 2016.